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Primer concentration for cDNA synthesis using GSP

cdna cloning GSP primer

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#1 PlantS_Student



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Posted 26 September 2012 - 09:59 AM

Hello Everyone,
I am trying to clone a 3.6 KB gene from the cDNA using a GSP. I am using the Superscript III enzyme from invitrogen and they specify using 2 pmol of GSP. So I made a 0.1 micromolar stock and used 1 microlitre for priming the cdna synthesis rxn. However the gene did not amplify. (I am going to use internal primers to check if atleast 1-2 kb of the gene amplified!) Does anyone have any experience or recommendations for using a particular primer concentration which works?
Thanks in Advance,

#2 bob1


    Thelymitra pulchella

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Posted 26 September 2012 - 12:40 PM

0.1 uM/l = 0.1 pM/ul...you didn't use enough primer.

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